Diabetes Research Group, Medical School, Institute of Cellular Medicine, North East Stem Cell Institute, Newcastle University, UK.
J Endocrinol. 2011 Nov;211(2):169-76. doi: 10.1530/JOE-11-0123. Epub 2011 Aug 18.
The source of new β-cells in adult human pancreas remains incompletely elucidated with recent studies on rodents providing evidence for neogenesis from progenitor cells in addition to self-replication. The aim of this study was to investigate the expression of pluripotency-associated stem cell markers in proliferative cultures derived from adult human pancreas. Human pancreatic tissue was obtained from deceased donors following ethical approval and relative consent. Islet-enriched fraction was separated from the retrieved organ by digestion and density gradient centrifugation. Dissociated cells were seeded in adherent culture forming proliferative 'islet survivor cells' (ISCs). These were characterised at fifth passage by RT-PCR, immunofluorescence staining, FACS, western blot and transfection studies with an OCT4 promoter-driven reporter. Nuclear expression of the pluripotency-associated stem cell marker complex OCT4/SOX2/NANOG was confirmed in ISCs. The phenotype constituted ∼8% of the overall population. OCT4 biosynthesis was confirmed by western blot and activation of an exogenous OCT4 promoter. Co-expression of pluripotency-associated markers has been confirmed in proliferative primary cells derived from adult human pancreas. Further studies are required to elucidate whether these cells possess functional stem cell characteristics and assess potential for differentiation into pancreatic cell lineages including new β-cells.
成体人胰腺内新β细胞的来源尚未完全阐明，最近对啮齿动物的研究为除了自我复制之外，祖细胞有新生的证据。本研究旨在探讨增殖培养物中多能性相关干细胞标志物在成体人胰腺中的表达。人类胰腺组织取自经过伦理批准和相对同意的已故供体。通过消化和密度梯度离心从回收的器官中分离胰岛富集部分。分离的细胞接种于贴壁培养中，形成增殖的“胰岛存活细胞”（ISCs）。在第五代通过 RT-PCR、免疫荧光染色、FACS、western blot 和 OCT4 启动子驱动的报告基因转染研究对这些细胞进行了特征分析。多能性相关干细胞标志物 OCT4/SOX2/NANOG 复合物的核表达在 ISCs 中得到了证实。该表型构成总体人群的约 8%。通过 western blot 证实了 OCT4 的生物合成，并激活了外源性 OCT4 启动子。增殖的原代细胞中已证实存在多能性相关标志物的共表达，这些细胞是否具有功能性干细胞特征，以及评估其分化为包括新β细胞在内的胰腺细胞谱系的潜力，还需要进一步研究。
